Jin Su-Eon, Kim Chong-Kook
College of Pharmacy, Seoul National University, Seoul, Republic of Korea; Department of Drug Development, College of Medicine, Inha University, Incheon, Republic of Korea.
College of Pharmacy, Seoul National University, Seoul, Republic of Korea; College of Pharmacy, Inje University, Gimhae, Gyeongnam, Republic of Korea.
Colloids Surf B Biointerfaces. 2014 Apr 1;116:582-90. doi: 10.1016/j.colsurfb.2014.01.053. Epub 2014 Feb 7.
Cationic lipid nanoparticles (cLNs) were modified to develop a gene delivery system for topical use via a dermal route. The cLNs were formulated using high pressure homogenization method and were composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), dioleoylphosphatidylethanolamine (DOPE), Tween 20, and tricaprin as a solid core (1:1:1:1.67, w/w). The prepared cLNs were nanoscale-sized (<100 nm) and were highly positive (51 mV). The cLN/DNA complexes demonstrated enhanced transfection potential in the cells at the optimal ratio without cytotoxic effects. To evaluate its efficacy in topical application, in vitro skin transfer of the cLN/DNA complexes was monitored using the measurement of the surface zeta potential of hairless mouse skin and validated using confocal microscopy of the sectioned skin. The in vivo delivery of plasmid DNA with the cLN formulation was examined using the relative expression levels of mRNA after non-invasive application with the cLN/DNA complexes on hair-removed dorsal skin of mice. The cLNs successfully transferred plasmid DNA to the skin, which was facilitated by the charge-mediated interaction between the cLN/DNA complexes and the skin. These results suggest the promising potential of cLNs as a topical gene delivery system for gene vaccine delivery and cutaneous gene therapy in preclinical and clinical applications.
阳离子脂质纳米粒（cLNs）经修饰后开发出一种用于经皮局部给药的基因递送系统。cLNs采用高压均质法制备，由1,2 - 二油酰基 - 3 - 三甲基铵丙烷（DOTAP）、二油酰磷脂酰乙醇胺（DOPE）、吐温20和作为固体核心的三癸酸甘油酯组成（1:1:1:1.67，w/w）。制备的cLNs为纳米级尺寸（<100 nm）且带强正电荷（51 mV）。cLN/DNA复合物在最佳比例下在细胞中表现出增强的转染潜力且无细胞毒性作用。为评估其局部应用的效果，通过测量无毛小鼠皮肤的表面zeta电位监测cLN/DNA复合物的体外皮肤转运，并使用切片皮肤的共聚焦显微镜进行验证。在小鼠脱毛背部皮肤无创应用cLN/DNA复合物后，通过mRNA的相对表达水平检测了cLN制剂对质粒DNA的体内递送情况。cLNs成功地将质粒DNA转运至皮肤，这得益于cLN/DNA复合物与皮肤之间的电荷介导相互作用。这些结果表明cLNs作为一种局部基因递送系统在临床前和临床应用中用于基因疫苗递送和皮肤基因治疗具有广阔的前景。
